Immunohistochemistry (IHC)
(Anti-CHAT antibody IHC staining of human small intestine. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody dilution 1:100.)
Rabbit anti-Mouse, Human CHAT Polyclonal Antibody | anti-CHAT antibody
Anti-CHAT Antibody (aa98-128) IHC-plus
IHC-P (1:100)
WB (1:1000)
Optimal dilution to be determined by the researcher
Usage:
This antibody does not work in IHC with samples fixed in Bouins. It has been tested with samples fixed in 10% neutral buffered formalin. HIER antigen retrieval with 10 mM sodium citrate is required.
Tissue Preparation:
Formalin fixation and embedding in paraffin wax.
Tissue Sectioning:
Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C.
Deparaffinization:
Wash dry slides in 3 changes of xylene - 5 minutes each @ RT
Rehydration:
Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT
Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT
Wash slides in 1 change of 80% alcohol - 3 minutes @ RT
Rinse slides in gentle running distilled water - 5 minutes @ RT
Antigen retrieval:
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) -1 minute @ RT
Immunostaining:
(Do not allow tissues to dry at any time during the staining procedure)
Apply a universal protein block - 20 minutes @ RT
Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT
Rinse slides in 1 X TBST - 1 minute @ RT
Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase streptavidin - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT
Wash slides in distilled water - 1 minute @ RT
Dehydrate:
(This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB)
Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT
Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT
Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT
Wash slides in 3 changes of xylene - 1 minute each @ RT
Apply coverslip
For long term storage store at -20 degree C.
Immunohistochemistry (IHC)
(Anti-CHAT antibody IHC staining of human small intestine. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody dilution 1:100.)
Immunohistochemistry (IHC)
(Anti-CHAT antibody IHC staining of human small intestine. Immunohistochemistry of formalin-fixed, paraffin-embedded tissue after heat-induced antigen retrieval. Antibody dilution 1:100.)
Immunofluorescence (IF)
(Confocal immunofluorescence of CHAT Antibody with NCI-H460 cell followed by Alexa Fluor 488-conjugated goat anti-rabbit lgG (green). DAPI was used to stain the cell nuclear (blue).)
Immunofluorescence (IF)
(Confocal immunofluorescence of CHAT Antibody with NCI-H460 cell followed by Alexa Fluor 488-conjugated goat anti-rabbit lgG (green). DAPI was used to stain the cell nuclear (blue).)
Western Blot (WB)
(Western blot of CHAT Antibody in NCI-H460 cell line lysates (35 ug/lane). CHAT (arrow) was detected using the purified antibody.)
Western Blot (WB)
(Western blot of CHAT Antibody in NCI-H460 cell line lysates (35 ug/lane). CHAT (arrow) was detected using the purified antibody.)
Western Blot (WB)
(Western blot of CHAT Antibody in mouse liver tissue lysates (35 ug/lane). CHAT (arrow) was detected using the purified antibody.)
Western Blot (WB)
(Western blot of CHAT Antibody in mouse liver tissue lysates (35 ug/lane). CHAT (arrow) was detected using the purified antibody.)
NCBI and Uniprot Product Information
NCBI Description
This gene encodes an enzyme which catalyzes the biosynthesis of the neurotransmitter acetylcholine. This gene product is a characteristic feature of cholinergic neurons, and changes in these neurons may explain some of the symptoms of Alzheimer's disease. Polymorphisms in this gene have been associated with Alzheimer's disease and mild cognitive impairment. Mutations in this gene are associated with congenital myasthenic syndrome associated with episodic apnea. Multiple transcript variants encoding different isoforms have been found for this gene, and some of these variants have been shown to encode more than one isoform. [provided by RefSeq, May 2010]
Uniprot Description
CHAT: an enzyme that catalyzes the reversible synthesis of acetylcholine (ACh) from acetyl CoA and choline at cholinergic synapses. Four alternative splice variants have been described.
Protein type: Acetyltransferase; EC 2.3.1.6; Lipid Metabolism - glycerophospholipid
Chromosomal Location of Human Ortholog: 10q11.2
Cellular Component: cytoplasm; cytosol; mitochondrion; neuron projection; nucleus
Molecular Function: choline binding; choline O-acetyltransferase activity
Biological Process: acetylcholine biosynthetic process; antral ovarian follicle growth; glycerophospholipid biosynthetic process; memory; neurotransmitter secretion; phosphatidylcholine biosynthetic process; phospholipid metabolic process; response to drug; response to ethanol; response to hypoxia; response to nutrient; synaptic transmission
Disease: Myasthenic Syndrome, Congenital, Associated With Episodic Apnea
Research Articles on CHAT
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Product Notes
The CHAT chat (Catalog #AAA249670) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-CHAT Antibody (aa98-128) IHC-plus reacts with Mouse, Human and may cross-react with other species as described in the data sheet. AAA Biotech's CHAT can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC) Paraffin, Immunofluorescence (IF), Western Blot (WB). IF (1:10 - 1:50) IHC-P (1:100) WB (1:1000) Optimal dilution to be determined by the researcher Usage: This antibody does not work in IHC with samples fixed in Bouins. It has been tested with samples fixed in 10% neutral buffered formalin. HIER antigen retrieval with 10 mM sodium citrate is required. Researchers should empirically determine the suitability of the CHAT chat for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CHAT, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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